US11150175B2ActiveUtilityA1
Light scattering detectors and methods for the same
Est. expiryJan 2, 2039(~12.4 yrs left)· nominal 20-yr term from priority
G01N 2021/513G01N 2021/4704G01N 2015/03G01N 30/86G01N 30/74G01N 21/51G01N 21/05G01N 15/1429G01N 21/53G01N 15/1434G01N 15/10G01N 2015/1022
91
PatentIndex Score
5
Cited by
28
References
20
Claims
Abstract
Methods for determining a radius of gyration of a particle in solution using a light scattering detector are provided. The method may include passing the solution through a flowpath in a sample cell, determining respective angular normalization factors for first and second angles of the detector, obtaining a first scattering intensity of the particle in solution at the first angle, obtaining a second scattering intensity of the particle in solution at the second angle, obtaining a 10° scattering intensity of the particle in solution at an angle of about 10°, determining a first particle scattering factor, determining a second particle scattering factor, plotting an angular dissymmetry plot, fitting a line to the angular dissymmetry plot, determining a slope of the line at a selected location on the line, determining the radius of gyration of the particle in solution from the slope of the line, and outputting the radius of gyration.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1. A method for determining a radius of gyration (Rg) of a particle in a solution using a light scattering detector, the method comprising:
passing the particle in the solution through a flowpath in a sample cell, wherein the flowpath has a centerline aligned with a beam of light directed through a portion of the detector;
determining an angular normalization factor (N θ1 ) for a first angle of the detector and an angular normalization factor (N θ2 ) of a second angle of the detector, wherein the first angle is about 90° relative to the centerline, and wherein the second angle is about 170° relative to the centerline;
obtaining a first scattering intensity (I θ1 ) of the particle in the solution at the first angle;
obtaining a second scattering intensity (I θ2 ) of the particle in the solution at the second angle;
obtaining a third scattering intensity (I 10 ) of the particle in the solution at a third angle of about 10° relative to the centerline;
determining a first particle scattering factor (P θ1 ) with the first scattering intensity (I θ1 ), the third scattering intensity (I 10 ), and the angular normalization factor (N θ1 ) for the first angle;
determining a second particle scattering factor (P θ2 ) with the second scattering intensity (I θ2 ), the 10° scattering intensity (I 10 ), and the angular normalization factor (N θ2 ) for the second angle;
plotting an angular dissymmetry plot, wherein the angular dissymmetry plot comprises the first particle scattering factor (P θ1 ) and the second particle scattering factor (P θ2 );
fitting a line to the angular dissymmetry plot;
determining a slope of the line at a selected location on the line;
determining the radius of gyration (Rg) of the particle in solution from the slope of the line; and
outputting the radius of gyration (Rg).
2. The method of claim 1 , wherein determining the angular normalization factor of the first and second angles of the detector comprises:
passing each of a plurality of known particles in the solution through the flowpath of the sample cell;
obtaining scattering intensity values for each of the plurality of known particles in the solution at the first angle, at the second angle, and at the third angle;
determining the angular normalization factor (N θ1 ) for the first angle with a plot of a ratio of the scattering intensity values of each of the plurality of known particles at the first angle to the scattering intensity values of each of the plurality of known particles at the third angle;
determining the angular normalization factor (N θ2 ) for the second angle with a plot of a ratio of the scattering intensity values of each of the plurality of known particles at the second angle to the scattering intensity values of each of the plurality of known particles at the third angle.
3. The method of claim 2 , wherein each of the plurality of known particles in the solution have a known molecular weight.
4. The method of claim 1 , wherein the first particle scattering factor (P θ1 ) is in the form
P
θ1
=
(
I
θ
1
I
10
)
N
θ
1
,
where:
I θ1 is the scattering intensity of the particle in the solution at the first angle;
I 10 is the scattering intensity of the particle in the solution at the third angle; and
N θ1 is the angular normalization factor for the first angle.
5. The method of claim 1 , wherein the second particle scattering factor (P θ2 ) is in the form
P
θ2
=
(
I
θ
2
I
10
)
N
θ2
,
wherein:
I θ2 is the scattering intensity of the particle in the solution at the second angle;
I 10 is the scattering intensity of the particle in the solution at the third angle; and
N θ2 is the angular normalization factor for the second angle.
6. The method of claim 1 , wherein plotting the angular dissymmetry plot comprises:
plotting a first point on a plane, the first point comprising a first coordinate and a second coordinate, wherein the first coordinate of the first point is the first particle scattering factor (P θ1 ), and wherein the second coordinate of the first point is in the form
μ
θ
1
2
=
(
4
π
n
0
sin
θ
1
2
λ
)
2
,
wherein:
n 0 is a refractive index of the solution;
θ 1 is the first angle; and
λ is a wavelength of the beam of light;
plotting a second point on the plane, the second point comprising a first coordinate and a second coordinate, wherein the first coordinate of the second point is the second particle scattering factor (P θ2 ), and wherein the second coordinate of the second point is in the form
μ
θ2
2
=
(
4
π
n
0
sin
θ
2
λ
)
2
,
wherein:
n 0 is a refractive index of the solution;
θ 2 is the second angle; and
λ is the wavelength of the beam of light.
7. The method of claim 1 , wherein fitting the line to the angular dissymmetry plot comprises a least squares fitting, and wherein the line comprises a polynomial degree of less than three.
8. A method for determining a radius of gyration (Rg) of a particle in a solution using a light scattering detector, the method comprising:
passing the particle in the solution through a flowpath in a sample cell, wherein the flowpath has a centerline aligned with a beam of light directed through a portion of the detector;
determining an angular normalization factor (N θ1 ) for a first angle of the detector, wherein the first angle is either about 90° or about 170° relative to the centerline;
obtaining a first scattering intensity (I θ1 ) of the particle in the solution at the first angle;
obtaining a 10° scattering intensity (I 10 ) of the particle in the solution at an angle of about 10° or less;
determining a first particle scattering factor (P θ1 ) with the first scattering intensity (I θ1 ), the 10° scattering intensity (I 10 ), and the angular normalization factor (N θ1 ) for the first angle;
plotting an angular dissymmetry plot, wherein the angular dissymmetry plot comprises the first particle scattering factor (P θ1 );
fitting a line to the angular dissymmetry plot;
determining a slope of the line at a selected location on the line;
determining the radius of gyration (Rg) of the particle in the solution from the slope of the line; and
outputting the radius of gyration.
9. The method of claim 8 , wherein determining the angular normalization factor (N θ1 ) for the first angle of the detector comprises:
passing each of a plurality of known particles in the solution through the flowpath of the sample cell;
obtaining scattering intensity values of each of the plurality of known particles in the solution at an angle of about 10° or less and at the first angle; and
determining the angular normalization factor (N θ1 ) for the first angle with a plot of a ratio of the scattering intensity values of each of the plurality of known particles at the first angle to the scattering intensity values of each of the plurality of known particles at an angle of about 10° or less with respect to a respective weight average molecular weight of each of the plurality of known particles in the solution.
10. The method of claim 9 , wherein each of the plurality of known particles in the solution have a known molecular weight.
11. The method of claim 8 , wherein the first particle scattering factor (P θ1 ) is in the form
P
θ1
=
(
I
θ
1
I
10
)
N
θ
1
,
where:
I θ1 is the scattering intensity of the particle in the solution at the first angle;
I 10 is the scattering intensity of the particle in the solution at an angle of about 10° or less; and
N θ1 is the angular normalization factor for the first angle.
12. The method of claim 8 , wherein plotting the angular dissymmetry plot comprises:
plotting a first point on a plane, the first point comprising a first coordinate and a second coordinate, wherein the first coordinate of the first point is the first particle scattering factor (P θ1 ), and wherein the second coordinate of the first point is in the form
μ
θ
1
2
=
(
4
π
n
0
sin
θ
1
2
λ
)
2
,
wherein:
n 0 is a refractive index of the solution;
θ 1 is the first angle; and
λ is a wavelength of the beam of light.
13. The method of claim 12 , wherein plotting the angular dissymmetry plot further comprises:
plotting a second point on the plane, the second point comprising a first coordinate and a second coordinate, wherein the first coordinate of the second point is the second particle scattering factor (P θ2 ), and wherein the second coordinate of the second point is in the form
μ
θ2
2
=
(
4
π
n
0
sin
θ2
2
λ
)
2
,
wherein:
n 0 is a refractive index of the solution;
θ 2 is the second angle; and
λ is the wavelength of the beam of light.
14. The method of claim 8 , wherein the line of the angular dissymmetry plot is a straight line.
15. The method of claim 8 , wherein the radius of gyration (Rg) of the particle in the solution is less than 10 nm.
16. The method of claim 8 , further comprising:
obtaining an angular normalization factor (N θ2 ) of a second angle of the detector, wherein the second angle is either about 90° or about 170° relative to the centerline, and wherein the second angle is different from the first angle;
obtaining a second scattering intensity (I θ2 ) of the particle in the solution at the second angle; and
determining a second particle scattering factor (P θ2 ) with the second scattering intensity (I θ2 ), the 10° scattering intensity (I 10 ), and the angular normalization factor (N θ2 ) for the second angle,
wherein the angular dissymmetry plot further comprises the second particle scattering factor (P θ2 ).
17. The method of claim 16 , wherein determining the angular normalization factor of the second angle of the detector comprises:
obtaining scattering intensity values of each of the plurality of known particles in the solution at the second angle; and
determining the angular normalization factor (N θ2 ) for the second angle with a plot of a ratio of the scattering intensity values of each of the plurality of known particles at the second angle to the scattering intensity values of each of the plurality of known particles at an angle of about 10° or less with respect to a respective weight average molecular weight of each of the plurality of known particles in the solution.
18. The method of claim 17 , wherein the second particle scattering factor (P θ2 ) is in the form
P
θ2
=
(
I
θ
2
I
10
)
N
θ2
,
wherein:
I θ2 is the scattering intensity of the particle in the solution at the second angle;
I 10 is the scattering intensity of the particle in the solution at an angle of about 10° or less; and
N θ2 is the angular normalization factor for the second angle.
19. The method of claim 16 , wherein the line of the angular dissymmetry plot is a curved line.
20. The method of claim 16 , wherein the radius of gyration (Rg) of the particle in the solution is less than 100 nm, optionally greater than 10 nm.Join the waitlist — get patent alerts
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